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  • SspI-HF®

    This enzyme has transitioned to an improved new buffer system. Visit NEBCutSmart.com for further details.
     
    The new and current Double Digest Finder and current Activity/Performance Chart for the CutSmart buffer system are available. The previous version of the Double Digest Finder, as well as the previous Version of Activity/Performance Chart that use the former buffer system, are still available for your convenience.

    High Fidelity Enzymes

    Product Update: A free vial of Gel Loading Dye, Purple (6x) is now included with all HF restriction enzymes. Please note there may be a slight delay for freezers and international products as inventory is switched over.
    cloned at neb recombinant engineered timesaver 5min incubation temp heat inactivation no
    Ssp-I-cutsite_1
    Catalog #SizeConcentrationPriceQtyAdd to Cart
    R3132S1,000 units20,000 units/ml$65.00Add to Cart
    R3132L5,000 units20,000 units/ml$260.00Add to Cart
    R3132M5,000 units100,000 units/ml$260.00Add to Cart
      
    Categories:
    High-Fidelity (HF®) Restriction Endonucleases,
    Restriction Endonucleases: S,
    Time-Saver™ Qualified Restriction Enzymes
    Applications:
    Restriction Enzyme Digestion

    Description

      
    High Fidelity (HF®) Restriction Enzymes have 100% activity in CutSmart™ Buffer; single-buffer simplicity means more straightforward and streamlined sample processing. HF enzymes also exhibit dramatically reduced star activity. HF enzymes are all Time-Saver™ qualified and can therefore cut substrate DNA in 5-15 with the flexibility to digest overnight without degradation to DNA. Engineered with performance in mind, HF restriction enzymes are fully active under a broader range of conditions, minimizing off-target products, while offering flexibility in experimental design.

    Product Source

    An E. coli strain that carries the cloned and modified (Y98F) SspI gene from Sphaerotilus species (ATCC 13925)

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    CutSmart Buffer-2010X

    Properties and Usage

    Unit Definition

    One unit is defined as the amount of enzyme required to digest 1 μg of λ DNA in 1 hour at 37°C in a total reaction volume of 50 μl.

    Reaction Conditions

    1X CutSmart™ Buffer
    Incubate at 37°C

    1X CutSmart™ Buffer:
    50 mM Potassium Acetate
    20 mM Tris-acetate
    10 mM Magnesium Acetate
    100 μg/ml BSA
    pH 7.9 @ 25°C

    Activity in NEBuffers

    NEBuffer 1.1: 25%
    NEBuffer 2.1: 100%
    NEBuffer 3.1: 10%
    CutSmart™ Buffer: 100%

    Diluent Compatibility

    Storage Temperature

    -20°C

    Storage Conditions

    10 mM Tris-HCl
    200 mM NaCl
    1 mM DTT
    0.1 mM EDTA
    200 μg/ml BSA
    50% Glycerol
    pH 7.4 @ 25°C

    Heat Inactivation

    No

    Methylation Sensitivity

    dam methylation: Not Sensitive
    dcm methylation: Not Sensitive
    CpG Methylation: Not Sensitive

    Quality Control

    Quality Control Assays

    The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
    • Exonuclease Activity (Radioactivity Release):
      The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
    • Ligation and Recutting (Terminal Integrity):
      After an over-digestion of DNA with a restriction endonuclease the percentage of the DNA fragments ligated with T4 DNA ligase and the percentage that can be recut are determined by agarose gel electrophoresis.
    • Non-Specific DNase Activity (16 hour):
      The product is tested for non-specific nuclease degradation in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.

    Supporting Documents

    Certificate of Analysis

    The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

    Specifications

    The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.

    Datacards

    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.
    1. When should I choose the HF version of the enzyme?
    2. When is star activity a concern?
    3. Why does the HF version of the enzyme have a different recommended buffer than the wild type enzyme?
    4. When should I choose the High Fidelity (HF) version of the enzyme?
    5. Can the change in buffer preference of the HF enzyme be advantageous?
    6. Will the HF enzyme produce elevated star activity when used in a buffer other than the one recommended?
    7. What does it mean to be Time-Saver™ qualified?
    8. How is the improvement in fidelity of HF restriction endonucleases quantitated?
    9. What is the Fidelity Index (FI)?
    10. How does the level of star activity of SspI-HF compare to SspI?
    11. What is the difference between SspI-HF and SspI?
    12. What is the specific activity of SspI-HF?
    13. Is there any difference in the methylation sensitivity between SspI-HF and SspI?
    14. Is there a difference in cutting close to the ends between SspI-HF and SspI?
    15. What does HF refer to following the name of a restriction enzyme?
    16. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
    17. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
    18. How can I access the old NEBuffer Activity Chart?
    19. How can I access the old Double Digest Finder?
    20. I tested your restriction enzyme on the substrate DNA recommended by NEB, and it appears to be active, however it does not digest my DNA. What could be the reason?
    21. My restriction enzyme was previously heat-inactivated, however, now it cannot be heat inactivated. Is there a reason for the change?

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